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The Histochemical Journal 31: 739–746, 1999.

© 1999 Kluwer Academic Publishers. Printed in the Netherlands.

Lectin binding pattern and proteoglycan distribution in human eccrine sweat glands

K. Sames1 , I. Moll2 , E.J.M. van Damme3 , W.J. Peumans3 & U. Schumacher1


1
Anatomical Institute, University of Hamburg, D-20246 Hamburg, Germany
2
Department of Dermatology, University of Hamburg, Germany
3
Laboratorium voor Fytopathologie en Plantenbescherming, Katholieke Universiteit Leuven, Belgium

Received 23 April 1999 and in revised form 29 July 1999

Summary

The distribution pattern of glycoconjugates in human eccrine sweat glands has been studied by the binding of newly discovered
lectins and by antibodies against a chondroitin sulphate proteoglycan and chondroitin sulphate glycosaminoglycans. Mannose-
specific lectins labelled large intracellular granules, part of which could be extended cisternae of the endoplasmic reticulum
or Golgi apparatus. In contrast, lectins specific for terminal mannose/glucose residues predominantly labelled basement mem-
branes and the glycocalyx. Lectins recognizing terminal N -acetylgalactosamine groups left most parts of the glands unstained,
but stained some dark cells intensely. These last cells were also intensively labelled by N -acetylglucosamine-specific and
by fucose-specific lectins. Sialic acid residues were preferentially located in luminal borders of secretory coils. No terminal
galactose residues were detected. All antibodies against chondroitin glycoconjugates stained large granules similar to those
revealed by the mannose-specific lectins in the secretory cells. The basement membrane is only stained by the proteoglycan
antibody and the chondroitin-6-sulphate antibody.
Thus, a complex composition of glycoconjugates exists not only in matrix elements but also in the cells of eccrine glands
of the human skin. A possible secretion of glycoconjugates is discussed.

Introduction histochemical methods, particularly in the dark cells of the


secretory coils and in the cell membranes of clear cells or
Two major portions of eccrine glands of the human skin can be intercellular canaliculi (Spicer et al. 1978, Tsukise et al.
distinguished, a ductular segment (dermal duct) and a secre- 1988). Using more specific techniques, however, individual
tory segment (secretory coil), where the duct cells show sim- glycosaminoglycans such as hyaluronic acid (Wang et al.
ilarity with the basal layer of the epidermis, while secretory 1992) and keratan sulphate were demonstrated in secretory
cells differ from epidermal cells in terms of cytokeratin types cells of eccrine glands (Sorell and Caterson 1990). The pres-
(Moll & Moll 1991, 1992). The dermal duct is lined by a ence of chondroitin sulphate was shown by an ultrahisto-
two-layered epithelium with a basal and an apical or luminal chemical technique in connection with basolateral but not
layer. In contrast, the secretory coil consists of a pseudo- with apical cell membranes of the secretory cells (Saga and
stratified epithelium containing myoepithelial cells, basally Takahashi 1993). However, chondroitin-type glycosamino-
located clear cells and dark cells which are predominantly glycans, the main extracellular proteoglycan-types, have
apically located. From a functional morphological point of hitherto not been demonstrated by antibodies of different
view, clear cells show a serous type of secretion and the dark specificities in normal eccrine glands of the human skin.
cells show certain characteristics of a mucous type secretion. The scope of the study reported in this paper, using a
The presence of different glycoconjugates, including both number of recently isolated lectins and antibodies to differ-
mucin type glycoproteins and glycosaminoglycans, has been ent chondroitin substances, was to classify the intracellular
demonstrated in the eccrine glands. Earlier investigations to mucous substances as well as glycoconjugate components of
demonstrate the existence of glycoproteins in the dark cells the glycocalyces and basal membranes in eccrine glands of
of the eccrine glands rested mainly on the periodic acid-schiff human skin.
(PAS) reaction (Spicer et al. 1978). Later, a number of lectins
were used to characterize defined carbohydrate residues of
glycoconjugates in all cell types of the secretory coils and Materials and methods
ducts of eccrine glands (Ookusa et al. 1983, Hazen-Martin
et al. 1986, Konick et al. 1988, Wollina et al. 1989, 1991, Normal skin areas adjacent to tumours from seven patients
Illana et al. 1997). were drawn from the files of the Dermatology Department,
The presence of glycosaminoglycans had been demon- University of Hamburg. The tissues were fixed in formalin,
strated earlier by pH-dependent as well as by metachromatic embedded in paraffin wax and processed for conventional
740 K. Sames et al.

light microscopy. The rehydrated sections were pretreated Table 2. Sources of proteoglycan antibodies.
with 0.1% trypsin in distilled water and incubated with
Source Glycosaminoglycan-specific Proteoglycan-specific
10 µg/ml biotinylated lectins (for source of the lectins, their
against against
abbreviations and nominal monosaccharide specificities, see
Table 1) for 1 h at room temperature. After careful wash- Seikagaku Chondroitin, unsulphated
Seikagaku Chondroitin-4-sulphate
ing in Tris-buffered saline (TBS; 50 mM Tris, 150 mM NaCl, Seikagaku Chondroitin-6-sulphate
1 mM MgCl2 , 1 mM CaCl2 , pH 7.4), biotin–avidin complexes Sigma Chondroitin sulphate
with phosphatase as the marker enzyme were used at a dilu- proteoglycan from
tion of 1 : 250 in lectin buffer for 30 min. New Fuchsin was chicken fibroblasts
used for visualization and the reaction was stopped with
4% buffered formalin. For nuclear counterstaining, Mayer’s
haematoxylin, diluted 1 : 1 v/v in distilled water was used. The Omission of chondroitinase treatment and/or the first anti-
slides were mounted in a water-soluble mounting medium bodies were used as negative controls. Rat articular cartilage
(crystal mount). and mast cells contain these glycosaminoglycans and were
Controls were performed by incubation with the lectin used as positive controls.
solutions containing their specific inhibitory monosaccha- Three different glands were evaluated in the tissue of
rides. each patient, stained with each of the lectins and antibodies,
For the immunostaining with the chondroitin/chondroitin respectively.
sulphate-specific antibodies rehydrated slides were digested
into chondroitinase ABC (Sigma) to unmask the specific
antigen epitope recognized by a particular antibody. Sec- Results
tions were incubated with 5U chondroitinase ABC in Tris
buffer, pH 8, for 4 h at 40 ◦ C in a moist chamber. Slides were Lectin histochemistry
washed with distilled water and phosphate-buffered saline
(PBS). This was followed by incubation with the first anti- N-acetylglucosamine (GlucNAc)-specific lectins (WGA,
body (dilution 1 : 200) overnight at 4 ◦ C in a moist chamber UDA, CMA) (for abbreviations of lectins, see Table 1)
(for specificity and source of first antibodies, see Table 2). An overview of an eccrine gland with secretory coils and der-
After a careful wash, samples were incubated with a second mal ducts is shown in Figure 1a. WGA stains the glands all
biotinylated antibody. Incubation with a PAP complex was over intensively. In the secretory coils, dark cells or the api-
followed by another washing step and incubation with an cal parts of dark cells (staining as an apical ring-shaped bor-
ABC complex. PAP was visualized by 3,30 -diaminobenzidine der) are preferentially stained by WGA with high intensity.
dihydrochloride and H2 O2 . After dehydration, the slides were However, a remarkable variation is observed. The clear cells
mounted in DEPEX. are also stained with high intensity and variability in part of
the glands (Figure 1b,c). The glycocalyx is not preferentially
labelled compared with other structures. The basement mem-
brane shows the same staining intensity as the glycocalyx.
Dermal duct cells are less intensively labelled by WGA
Table 1. Sources of lectins. than cells of the secretory portion. The apical cell layer shows
Source Abbreviation Nominal sugar Supplier medium staining intensities. The basal layer remains pale
specificity and a ring-shaped apical border is stained occasionally and
Triticum aestivum WGA GlucNAc Sigma
is not restricted to the glycocalyx. The basement membrane
Urtica dioica UDA GlucNAc + is labelled in some parts with the same intensity (Figure 1b).
Chelidonium majus CMA GlucNAc + The UDA and CMA staining patterns are similar to the
Glycine max SBA GalNAc Sigma staining pattern of WGA. With UDA, an intensive apical
Iris reticulata IRA GalNAc + staining is less frequent in the secretory coils compared to
Colchicum autumnale CAA GalNAc +
Aegopodium podagraria APA GalNAc +
the other lectins of this group. CMA labels clear cells more
Amaranthus caudatus ACA Galactose + intensively than WGA (compare Figure 1a with 1c).
Allium ursinum AUA Mannose + Treatment with the specific monosaccharide of this group
Listera ovata LOA Mannose + of lectins did not totally extinguish the reaction. This unspe-
Galanthus nivalis GNA Mannose + cific reaction is sometimes intense and resembles the reaction
Hippeastrum hybrid HHA Mannose +
Canavalia ensiformis Con-A Mannose/Glucose Sigma
of chondroitin sulphates.
Lens culinaris LCA Mannose/Glucose Sigma
Ulex europeus UEA-I Fucose Sigma N-acetylgalactosamine (GalNAc)-specific lectins
Sambucus nigra SNA I Sialic acid + (SBA, IRA, CAA, APA)
+ = inhouse supply (E.J.M. van Damme, W.J. Peumans); see Brooks In general, the staining intensity ranges from low to medium
et al. (1987) and Van Damme et al. (1998). or is even lacking completely (Figure 2a,b). Dark cells or
GlucNAc = N-acetylglucosamine, GalNAc = N-acetylgalatosamine. apical parts of dark cells, however, are intensely labelled in
Lectin binding and proteoglycan distribution in human eccrine sweat glands 741

Figure 2. Staining pattern of GalNAc-specific lectins in eccrine sweat


glands. (a) IRA; secretory coil and transition to the sweat duct; fine gran-
ular staining of secretory vesicles in dark cells (arrow); 410× (b) SBA;
secretory coil; unstained cells and medium intensity of the basement
membrane; 410×.

the staining of intracellular structures and is hence difficult


to identify from intracellular structures (Figure 2a).
IRA labels the basement membrane of secretory coils with
the same intensity as it produces in dark cells (Figure 2b).

Galactose (Gal)-specific lectin (ACA)


The reaction with this lectin is negative.

Figure 1. Staining pattern of GlucNAc-specific lectins in eccrine sweat Mannose (Man)-specific lectins (AMA, LOA, GNA, HHA)
glands. (a) CMA; intensive staining of dark cells and clear cells; 130× The cytoplasmic staining of dark cells ranges from very
(b) WGA; sweat ducts (secretory tubule at the left); arrow points to api- high to low in different cells of the secretory coil (compare
cal densification; 410× (c) WGA; cross-section of secretory coil: pro- Figure 3a,b). The staining of clear cells follows a similar pat-
nounced staining of dark cells; 410×. tern. Both types of cells often show intensely stained large
granules even if cytoplasmic staining is pale (Figure 3b). Both
some parts of a number of secretory coils (Figure 2a). The the glycocalyx and the basement membrane show almost the
lectin binding intensity of clear cells ranges from unlabelled same staining pattern as compared with the cytoplasm and
to medium. Both glycocalyx and basement membrane show hence they are hardly discernable.
low to medium labelling intensities. The dermal duct cells react with medium to low intensity.
In dermal ducts, the apical and basal cells show medium In the apical cytoplasm of the apical cells, high reaction inten-
reactivity. The glycocalyx is pronounced in parts of some sities can be observed occasionally. The basement membrane
tubules. The staining of the basement membrane merges with is intensely stained (Figure 3a).
742 K. Sames et al.

Figure 3. Staining pattern of Man-specific lectins in eccrine sweat


glands. (a) AMA; secretory coil and dermal duct (lower medial part); Figure 4. Staining pattern of Man/Glu-specific lectins in eccrine sweat
410× (b) GNA; labelling of granules (arrows) discernable from cell glands; intensive reaction of the basement membranes. (a) Con A; secre-
nuclei (stained by the counterstain, arrowheads); 410×. tory coil; 410×. (b) Con A; secretory coil; 410×.

Mannose/glucose (Man/Glu)-specific lectins (Con A, LCA)


These lectins preferentially react with the basement mem-
brane of the glands (Figure 4a,b). The cytoplasm of dark
cells shows medium intensities in most parts of the major-
ity of the glands. However, in some parts of a number of
secretory coils, dark cells are stained intensely. Clear cells
show the same staining pattern (Figure 4a). The glycocalyx
is pronounced, however, with a lower labelling intensity as
compared to the basement membrane.
In the dermal ducts, both apical and basal cells show
medium intensities. The basement membrane is intensely
labelled, the glycocalyx reacts occasionally.

Fucose (Fuc)-specific lectin (UEA-I)


In some glands, a part of the dark cells or the apical cytoplasm
of dark cells show pronounced lectin reactivity of medium
to high intensities. Lectin binding in the cytoplasm of the Figure 5. Staining pattern of Fuc-specific lectins in eccrine sweat glands;
clear cells is weak. The glycocalyx is not discernable from UEA-I; exclusive staining of dark cells; 260×.
the apical cytoplasm. The basal membrane shows medium
labelling intensities (Figure 5). Sialic acid-specific lectin (SNA-I)
In the dermal duct the cytoplasm of the apical and basal Overall staining of the secretory coil cells is weak. In the
cells is pale or remains unstained. The glycocalyx stains with glands, a wide apical rim, presumably representing the api-
low to medium intensity. The basal membrane is weakly cal parts of the dark cells, shows an intensive reaction in a
labelled or even unstained. number of cross-sections. The basal cytoplasm of clear cells
Lectin binding and proteoglycan distribution in human eccrine sweat glands 743

shows low to medium reactivity or no reaction at all. The Proteoglycan and glycosaminoglycan histochemistry
glycocalyx staining is also of low to medium intensity. The
basal membrane stains with the same intensity as the glyco- Chondroitin sulphate proteoglycan antibody
calyx. Two specimens exhibited low cytoplasmic labelling of The majority of cells are unstained (Figure 6a). However,
all cells. large granules in the apical cytoplasm of some dark cells
In the ducts, apical and basal cells stain with low to medium react for chondroitin sulphate proteoglycan (Figure 6b). Fur-
intensity. In contrast to the secretory coils, the glycoca- thermore, an intensive reaction with other irregularly dis-
lyx shows higher labelling intensities as compared with the tributed and shaped intracellular granules or vacuoles exists.
cytoplasm. The basement membrane also labels intensively
occasionally.
In general, reactions vary in the secretory coils between
different cells and structures of the same type, between differ-
ent glands and between different sections of the same gland.
Weakly staining structures as well as the cell borders are often
hard to discriminate. Therefore, only the intense reactions
found in the different structures are given in Table 3. In the
dermal ducts, staining of structures is more homogenous and
is similar in different cross-sections. Therefore, high inten-
sive, low intensive and negative reactions can be assessed
(Table 4).

Table 3. Positive lectin staining in structures of the secretory


coils.
Structure BM Glycc APB Ccell Dcell Gran
specificity
GlcNac + + +
GalNac + +
Man + + +
Man/gluc + +
Fuc + +
Sia + +

+ = intensive staining (intensity higher than in other struc-


tures) is found frequently in these structures.
BM = basement membrane, Glycc = glycocalyx, APB = apical
boder, Ccell = clear cell, Dcell = dark cell, Gran = special
(large) granule.

Table 4. Lectin staining in structures of the dermal


ducts.
Structure BM Glycc APB Bcell Acell
specificity
GlcNac ++ ++ ++ + +
GalNac + ++ + + +
Man ++ + + + +
Man/gluc + + + + +
Gal − − − − −
Fuc +/− + + +/− +/−
Sia ++ ++ + + +

++ = intensive reaction, + = low to medium reaction


intensity, − = no reaction, +/− = the structure shows Figure 6. Chondroitin sulphate proteoglycan staining. (a) Secretory coil
positive and negative reacting parts. with main reactivity in the basal membranes; 410×. (b) Cells in the
BM = basement membrane, Glycc = glycocalyx, secretory coil with patchy apical reaction; 260×. (c) Nearly complete
APB = apical boder, Bcell = basal cell, Acell = apical staining of the cytoplasm, a reaction pattern observed only occasionally;
cell. 410×.
744 K. Sames et al.

Confluent small granules form large patches which some-


times fill the entire cytoplasmic space (Figure 6c). It is impos-
sible to decide whether the cells are dark or clear ones. The
glycocalyx is only stained in single sections through the secre-
tory coils or dermal ducts. Basement membranes are always
well stained and are the only stained structures in some parts
of the secretory coil or dermal duct.
In the dermal ducts cell membranes and basement mem-
branes are stained as well as cytoplasmic granules. The gly-
cocalyx behaves as in the secretory coils.

Chondroitin-6-sulphate
With the chondroitin-6-sulphate antibody, the reaction is very
similar to that against the chondroitin sulphate proteoglycan.
In dark cells, the apical granules are less regularly distributed
as compared with the chondroitin sulphate proteoglycan reac-
tion. Staining of the basement membrane is fragmentary.
Staining of the total cytoplasm is observed more frequently
as compared to the proteoglycan (Figure 7a,b).
The staining pattern of the dermal ducts is similar to
that obtained with the chondroitin sulphate proteoglycan
antibody.

Figure 8. Unsulphated chondroitin staining. (a) Secretory coil and sweat


duct; 260×. (b) Cross-section of another coil; 260×.

Chondroitin-4-sulphate
The reaction of the secretory coil and dermal duct is simi-
lar to that produced by the chondroitin 6-sulphate antibody.
However, the basement membrane only sporadically shows
a weak fragmentary immunoreactivity.

Unsulphated chondroitin
Cells are labelled as with the chondroitin sulphates. From the
immunostaining, clear cells are indistinguishable from the
dark cells. Staining of the total cytoplasm is more frequent as
compared to the chondroitin sulphates. Stained parts of the
glycocalyx also are visible more frequently. The basement
membrane remains unstained (Figure 8).
An overview of proteoglycan and glycosaminoglycan spe-
cific reactions is given in Table 5.

Discussion

The lectins UDA, CMA (GlucNAc-specific), IRA, CAA,


APA (GalNAc-specific), ACA (Gal-specific), AUA, LOA,
GNA, HHA (Man-specific), LCA (Man/Gluc-specific) and
SNA-I (sialic acid-specific) have so far not been used for
Figure 7. Chondroitin-6-sulphate staining; staining pattern very simi- investigating the histochemistry of sweat glands. These new
lar to that of the chondroitin sulphate proteoglycan; basal membrane
less intensively labelled. (a) Patchy cytoplasmic staining of cells in lectins share monosaccharide specificities with those previ-
the tubules; 260×. (b) Cross-section with nearly complete cytoplasmic ously used by other groups on eccrine glands with the excep-
staining; 410×. tion of the Man-specific ones. So far, only lectins which bind
Lectin binding and proteoglycan distribution in human eccrine sweat glands 745

Table 5. Positive and negative proteoglycan/glycosaminoglycan stain- very similar to, and may be identical with, mucins of other
ing in different structures of sweat glands. epithelial origin like those contained in mucus of the gastro-
intestinal or respiratory tracts. They are stored in intracellular
Structure BM Glycc APB Ccell/Dcell Gran Agran
specificity granules and secreted to protect epithelial surfaces (Strous &
Dekker 1992).
CSPG + −/+ − +/− + +
C-6-S +/− −/+ − +/− + +
In the apical cell layer of dermal ducts, the reaction patterns
C-4-S −/+ −/+ − +/− + + of the new GlcNAc-, GalNAc-, Man/Glu- and sialic acid-
Ch − +/− − + + + specific lectins and earlier results obtained with other lectins
of the same carbohydrate specificity are in good agreement
+ = positive reaction, − = negative reaction, +/− positive reacting
parts dominate over negative ones and −/+ vice versa. (Hazen-Martin et al. 1986, Tsukise et al. 1988, Wollina et al.
BM = basement membrane, Glycc = glycocalyx, APB = apical boder, 1989, 1991).
Ccell/Dcell = clear cells/dark cells (not discernable), Gran = special Some lectins produce a small apical lining of otherwise
(large) granules, Agran = apical situated large granules, CSPG = chon- unstained cells, presumably representing the glycocalyx.
droitin sulphate proteoglycan, C-6-S = chondroitin-6-sulphate, C-4-S
Man/Gluc-specific lectins (Con A, LCA) react with this gly-
= chondroitin-4-sulphate, Ch = unsulphated chondroitin,.
cocalyx in the secretory coil and GlcNAc-specific, GalNAc-
specific, Man-specific as well as sialic acid-specific ones react
with it in dermal ducts.
to Man and at the same time to Glu-like Con A and LCA Gibinbski et al. (1978), using electron microscopy, could
have been used for the lectin histochemistry of human sweat only demonstrate glycocalyx following sweating. This could
glands. The new Man-specific lectins label irregularly-stained indicate that substances coating cell surfaces are stored in the
large intracytoplasmic granules in both dark and clear cells, cells and secreted with the sweat. Tamaki et al. (1985) found
which are only sporadically labelled by the other lectins. a reaction restricted to the cell surfaces in eccrine glands using
Such granules are similar in appearance to those revealed FITC-labelled PNA and WGA but we have no corresponding
by proteoglycan/glycosaminoglycan immunohistochemistry. results using the new lectin ACA (which is GalNAc-specific
The large granules are different from the small apically like PNA) and WGA.
located secretory vesicles often visible by lectin staining. Part Our results clearly demonstrate the presence of unsul-
of these large granules could represent extended cisternae of phated, 4-sulphated, and 6-sulphated chondroitin as well as
the endoplasmic reticulum or Golgi apparatus (Schumacher a chondroitin sulphate proteoglycan in normal eccrine glands
et al. 1998), since Man residues are added to the protein chain of human skin. It is well known that chondroitins/chondroitin
of glycoproteins and of the corneal type of keratan sulphate proteoglycans represent normal constituents of basement
in the endoplasmic reticulum. membranes and glycocalyces. However, these glycoconju-
There is good agreement between our results and those of gates are not restricted to the extracellular and pericellular
others concerning lectin binding of the dark cells. GlucNAc- compartment of the sweat glands. We found them also within
specific (Tsukise et al. 1988, Meyer and Bartels 1989, Wollina epithelial cells.
et al. 1991, Illana et al. 1997), GalNAc-specific (Hazen- Mori et al. (1994) also have used anti-chondroitin-4-
Martin et al. 1986, Tsukise et al. 1988, Meyer and Bartels sulphate and anti-chondroitin-6-sulphate antibodies. In skin
1989, Tsubura et al. 1992, Illana et al. 1997), and sialic acid- specimens containing mixed tumours of the skin, normal
specific (Hazen-Martin et al. 1986) lectin binding as well as eccrine glands showed no reactivity. Hence the expression
the negative Gal-specific reaction (Tsubura et al. 1992) are of these substances by the epithelial tumour cells of eccrine
corroborated by the new lectins. gland origin have been interpreted as evidence for chondroid
In clear cells, intense binding of the new lectins UDA and changes in the tumour tissue. This observation is in contrast
CMA is in accordance with strong WGA staining (Terui et al. to our results and those of Sorrel and Caterson (1990) reveal-
1986), which we have used as a positive control for the group ing a positive reaction with anti-keratan sulphate antibodies.
of GlucNAc-specific lectins. With this group, we found, espe- This apparent contradiction may be explained by the different
cially in sweat glands, a positive reaction in the controls inhib- sensitivities of the methods used.
ited by the specific monosaccharide. Since this unspecific Chondroitin substances can be found in large granules or
reaction is similar to those of chondroitin and chondroitin vacuoles, which in part are also demonstrated by Sorrel and
sulphates, it can be explained by a reaction of the lectins Caterson (1990) using keratan sulphate antibodies. It is not
with these substances. Schumacher et al. (1992) have found clear if different chondroitin sulphate compounds are located
previously a positive reaction of WGA with chondroitin-4- in the same granules or in separate ones. The proteoglycan
sulphate. demonstrated by our antibody reacts in a pattern very similar
In the secretory coils, the intracellular lectin-reactivity may to that of the chondroitin-6-sulphate antibody. Chondroitin-
be caused by mucin-type glycoproteins (most of them are 6-sulphate may, therefore, be its main side chain.
O-linked oligosaccharides), especially those containing sialic Granules or vacuoles share the feature of proteoglycan
acid, GalNac, GlucNac or Fuc. The specific lectins for these immunoreactivity and binding of lectins of the Man-specific
monosaccharides bind to secretory cells and duct cells in our group. Again it remains unclear whether these granules con-
material. Such glycoproteins of eccrine glands of the skin are tain a mixture of all these glycoconjugates or whether they
746 K. Sames et al.

can be separated by specific glycoprotein, proteoglycan or Moll I, Moll R (1992) Changes of expression of intermediate filament
glycosaminoglycan content. It is interesting that different proteins during ontogenesis of eccrine sweat glands. J Invest Dermatol
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Mori M, Shrestha P, Sakamoto F, Yang LJ, Qin C, Tsujimura T (1994)
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substrates seem to be contained in special cells. This could tumour of the skin: immunohistochemical evaluation of bone morpho-
indicate a specialized secretion or a special synthetic status genetic protein, glycosaminoglycans, keratin, vimentin and neuronal
of particular cells. markers. Arch Dermatol Res 286: 285–292.
Our findings demonstrate an intensive GlcNac-, GalNac-, Ookusa Y, Takata K, Nagashima M, Hirano H (1983) Distribution of gly-
coconjugates in normal human skin using biotinyl lectins and avidin-
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